Development of an in vitro model for acute lymphoblastic leukemia
Author
Kiørboe, Katrine Juhlin
Term
4. term
Publication year
2022
Abstract
Acute lymphoblastic leukemia (ALL) is the most common childhood cancer, but ALL cells die rapidly outside the body, making laboratory studies difficult. This project aimed to develop an in vitro model that mimics the bone marrow microenvironment to better sustain ALL cells. The model combined co-culture of ALL cells with primary mesenchymal stromal cells on a coating of collagen, laminin, and fibronectin, alongside testing culture media with added cytokines (FLT-3, IL-2, IL-3, IL-7, SCF) and human platelet lysate (hPL). Stromal cell expression of the stem cell marker nestin was examined under normoxia and hypoxia in mono- and co-culture, and ALL cell survival was assessed by automated microscopy; AraC sensitivity was also evaluated in the model. Results showed that cytokines and hPL significantly increased ALL cell survival, all stromal cell types expressed nestin (strongest in co-culture), and co-culture and the developed model improved survival in both normoxia and hypoxia compared with monoculture. Overall, the study indicates that stromal support, ECM components, cytokines/growth factors, and hypoxia jointly help maintain ALL cell survival in vitro, providing a more realistic platform for future studies and drug testing.
Akut lymfoblastisk leukæmi (ALL) er den hyppigste børnekræft, men ALL-celler dør hurtigt uden for kroppen, hvilket gør laboratoriestudier udfordrende. Dette projekt havde til formål at udvikle en in vitro-model, der efterligner knoglemarvens mikromiljø for bedre at understøtte ALL-celler. Modellen bestod af co-kultur mellem ALL-celler og primære mesenkymale stromale celler på en belægning af kollagen, laminin og fibronektin, med afprøvning af forskellige medier, tilsatte cytokiner (FLT-3, IL-2, IL-3, IL-7, SCF) og human blodpladelysat (hPL). Stromale cellers udtryk af stamcellemarkøren nestin blev undersøgt i normoksi og hypoksi i både mono- og co-kultur, og ALL-cellers overlevelse blev vurderet med automatiseret mikroskopi; AraC-følsomhed blev også evalueret i modellen. Resultaterne viste, at cytokiner og hPL signifikant øgede ALL-cellers overlevelse, at alle stromale celletyper udtrykte nestin (stærkest i co-kultur), og at co-kultur og den udviklede model forbedrede overlevelsen i både normoksi og hypoksi sammenlignet med monokultur. Samlet peger studiet på, at kombinationen af stromal støtte, ECM-komponenter, cytokiner/vækstfaktorer og hypoksi er central for at fastholde ALL-cellers overlevelse in vitro og giver et mere realistisk grundlag for fremtidige studier og lægemiddeltest.
[This apstract has been generated with the help of AI directly from the project full text]